Mechanism of resveratrol in alleviating oxygen-glucose deprivation/reperfusion injury in primary cortical neurons
XUE Linmin
GUAN Xin
XU Weijie
YANG Liqiang
Abstract：Objective To investigate the mechanism by which resveratrol(Res)attenuates oxygen-glucose depriva-tion/reperfusion(OGD/R)-induced injury in primary cortical neurons based on silent information regulator1/forkhead box O1(SIRT1/FOXO1)signaling pathway.Methods We cultured primary cortical neurons in vitro and established the OGD/R injury models.After intervening with different concentrations of Res,neuronal viability was detected using the CCK-8 assay to screen the optimal experimental concentration of Res.The OGD/R-induced neuronal models were random-ly divided into three groups:OGD/R group,OGD/R+Res group,and OGD/R+Res+SIRT1 inhibitor EX527 group(OGD/R+Res+EX527 group),respectively;meanwhile,some normal primary cortical neurons were assigned as the control group(Norm group).We treated the OGD/R+Res group with the optimal concentration of Res,while the OGD/R+Res+EX527 group was administered the optimal concentration of Res combined with 25 μmol/L EX527 for 24-hour incubation.Corti-cal neuronal viability was determined via the CCK-8 assay.Quantitative real-time polymerase chain reaction(qRT-PCR)was applied to detect the mRNA expression levels of silent information regulator 1(SIRT1)and autophagy-associated gene microtubule-associated protein light chain 3(LC3).We performed Western blotting to measure the protein expression lev-els of SIRT1/forkhead box O1(FOXO1)signaling pathway-related proteins and autophagy-associated marker proteins(LC3-Ⅱ/LC3-Ⅰ ratio,p62),and immunofluorescence staining was used to observe the formation of autophagosomes(re-flected by LC3-positive cells).Results Based on the neuronal viability results,we selected 20 μmol/L as the optimal experimental concentration of Res.Compared with the Norm group,the mRNA expression levels of IRT1 and LC3 in the OGD/R group were slightly higher,but no statistically significant differences were found(both P>0.05).Compared with the OGD/R group,the mRNA expression levels of SIRT1 and LC3 increased in the OGD/R+Res group(both P<0.05),whereas the OGD/R+Res+EX527 group exhibited significantly lower mRNA expression levels of these two genes than those in the OGD/R+Res group(both P<0.05).Compared with the Norm group,the OGD/R group had significantly higher pro-tein expression levels of SIRT1 and FOXO1 as well as an elevated LC3-Ⅱ/LC3-Ⅰ ratio(all P<0.05),while the protein expression of p62 significantly decreased(P<0.05).Compared with the OGD/R group,the SIRT1 protein expression and LC3-Ⅱ/LC3-Ⅰ ratio were higher in the OGD/R+Res group(both P<0.05),and the protein expression levels of FOXO1 and p62 were significantly down-regulated(both P<0.05).Compared with the OGD/R+Res group,the OGD/R+Res+EX527 group showed significantly lower SIRT1 protein expression and LC3-Ⅱ/LC3-Ⅰ ratio(both P<0.05),along with significantly higher protein expression levels of FOXO1 and p62(both P<0.05).Additionally,a significantly higher pro-portion of LC3-positive cells was found in the OGD/R group than in the Norm group(P<0.05).We observed a significant-ly higher proportion of LC3-positive cells in the OGD/R+Res group than in the OGD/R group(P<0.05),while the propor-tion of LC3-positive cells in the OGD/R+Res+EX527 group was significantly lower than that in the OGD/R+Res group(P<0.05).Conclusion Res may alleviate OGD/R-induced primary cortical neuronal damage by promoting autophagy through activating the SIRT1/FOXO1 signaling pathway.
Keywords：resveratrolprimary cortical neuronal injuryautophagysilent information regulator1/forkhead box O1 signaling pathwayoxygen-glucose deprivation/reperfusion
Publication Date：2026-02-25
Online Publishing Date：2026-03-19(First online date of this platform, not the publication date of the document)
Pages：6( 34-38,43 )